Microbial composition

ABSTRACT

A synergistic microbicidal composition comprising: (a) at least one microbicide selected from the group consisting of a monosubstituted phenol and an isopropyl methyl phenol; and (b) at least one microbicide selected from the group consisting of substituted cyclohexyl propyl-1,3-diols, propen-2yl-methyl cyclohexanols and menthadiene alcohols.

This invention relates to a synergistic combination of selectedmicrobicides having greater activity than would be observed for theindividual microbicides.

It particularly relates to an microbiocidal composition for personalcleaning, oral care or hard surface cleaning or industrial andinstitutional cleaning applications.

In some cases, commercial microbicides cannot provide effective controlof microorganisms, even at high use concentrations, due to weak activityagainst certain types of microorganisms, e.g., those resistant to somemicrobicides, or due to aggressive environmental conditions.

For example, sanitising and disinfecting soap compositions comprisingchlorine-based antimicrobial agent such as triclosan are known. Suchcompositions require a rather long contact time to provide efficaciousantimicrobial action. In practice, users, in particular children, do notspend a long time on cleansing and as a result cleaning with suchcompositions does not provide adequate prevention from surface ortopical infection or adequate protection against diseases. The user, inspite of cleaning his hands, is generally likely to end up withrelatively inadequate bacterial removal from his skin. Therefore, he maycause contamination of further animate and/or inanimate surfaces andcontribute to the spreading of pathogens and consequent diseases. Usersin general and children in particular who wash contaminated hands beforemeals with slow-acting antimicrobial compositions for relatively shorttime are at risk of contracting diseases.

Similarly in the area of hard surface cleaning, e.g. cleaning of floors,table tops or utensils, the antimicrobial in the compositions are incontact with the substrate for less than a few minutes after which thesurface is either wiped off or rinsed with water. These short timescales of cleaning action are ineffective in providing the desiredbenefit since most known antimicrobials commonly used in such productstake many minutes to hours to provide the desired kill of microbes.

Therefore, there is a need of providing a composition that—uponapplication—provides relatively more efficacious antimicrobial actionduring a relatively short cleaning period, preferably about 30 secondsor less.

Combinations of different microbicides are sometimes used to provideoverall control of microorganisms in a particular end use environment.For example, WO2010/046238 discloses combinations of thymol andterpineol. However, there is a need for additional combinations ofmicrobicides having enhanced fast-acting activity against variousstrains of microorganisms to provide effective control of themicroorganisms. Moreover, there is a need for combinations containinglower levels of individual microbicides for safety, environmental,aesthetic and economic benefit. The problem addressed by this inventionis to provide such additional combinations of microbicides.

STATEMENT OF THE INVENTION

The present invention is directed to a synergistic microbicidalcomposition comprising: (a) at least one microbicide selected from thegroup consisting of 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol,2-menthyl-5-(prop-1-en-2-yl)cyclohexanol and5-methyl-2-(prop-1-en-2-yl)cyclohexanol; and (b) at least onemicrobicide selected from the group consisting of5-isopropyl-2-methylphenol; 3-isopropyl-5-methylphenol,4-isopropyl-3-methylphenol, (E)-2-(prop-1-enyl)phenol, 4-propylphenol,2-tert-butylphenol, 2-sec-butylphenol, 2-n-propylphenol,3-n-propylphenol, 4-n-butylphenol, 4-pentylphenol, 4-sec-butylphenol,and 3-tert-butylphenol.

The present invention is further directed to a synergistic microbicidalcomposition comprising: (a) 2-(trans-4-ethylcyclohexyl)propane-1,3-diolor 2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol; and (b) at least onemicrobicide selected from the group consisting of 4-propylphenol,2-n-propylphenol and 3-n-propylphenol.

The present invention is directed to a synergistic microbicidalcomposition comprising: (a) at least one microbicide selected from thegroup consisting of 2-methyl-5-(prop-1-en-2-yl)cyclohexanol and5-methyl-2-prop-1-en-2-yl)cyclohexanol; and (b) at least one microbicideselected from the group consisting of 5-isopropyl-2-methylphenol,4-isopropyl-3-methylphenol, (E)-2-(prop-1-enyl)phenol, 4-propylphenol,2-tert-butylphenol, 2-sec-butylphenol, 2-n-propylphenol,3-n-propylphenol, 4-n-butylphenol, 4-sec-butylphenol and3-tert-butylphenol.

The microbiocide 5-methyl-2-(prop-1-en-2-yl)cyclohexanol is preferably(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol.

The present invention is further directed to a synergistic microbicidalcomposition comprising: (a) 2-(trans-4-ethylcyclohexyl)propane-1,3-diol,and (b) at least one microbicide selected from the group consisting of4-propylphenol, 2-n-propylphenol and 3-n-propylphenol.

The present invention is further directed to a synergistic microbicidalcomposition comprising: (a)2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol; and (b)3-n-propylphenol.

The present invention is further directed to a synergistic microbicidalcomposition comprising: (a) 2-methyl-(prop-1-en-2-yl)cyclohex-2-enol;and (b) at least one microbicide selected from the group consisting of5-isopropyl-2-methylphenol; 3-isopropyl-5-methylphenol,4-isopropyl-3-methylphenol, (E)-2-(prop-1-enyl)phenol, 4-propylphenol,2-tert-butylphenol, 2-sec-butylphenol, 2-n-propylphenol,3-n-propylphenol, 4-n-butylphenol, 4-pentylphenol, 4-sec-butylphenol,and 3-tert-butylphenol.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the following terms have the designated definitions,unless the context clearly indicates otherwise. The term “microbicide”refers to a compound capable of killing, inhibiting the growth of orcontrolling the growth of microorganisms at a locus; microbicidesinclude bactericides, fungicides and algaecides. The term“microorganism” includes, for example, fungi (such as yeast and mold),bacteria and algae. The following abbreviations are used throughout thespecification: mL=milliliter, ATCC=American Type Culture Collection, andMBC=minimum biocidal concentration. Unless otherwise specified,temperatures are in degrees centigrade (° C.), and references topercentages are by weight (wt %).

The compositions of the present invention unexpectedly have been foundto provide enhanced microbicidal efficacy at a combined activeingredient level lower than that of the individual microbicides.Additional microbicides beyond those listed in the claims may be presentin the composition.

The present invention provides for a synergistic antimicrobialcomposition comprising a phenolic compound and a antimicrobial alcoholpreferably a terpene alcohol. The phenolic compound is preferablyselected from the class consisting of isopropyl methyl phenols andmonosubstituted phenols. The antimicrobial alcohol is preferablyselected from the class consisting of menthadiene alcohols,propen-2yl-methyl-cyclohexanol and substituted cyclohexylpropyl-1,3-diols.

The compounds claimed as combinations in the present invention and theclass they belong to are given below:

Class of propen-2yl-methyl Cyclohexanols:

-   2-methyl-5-(prop-1-en-2-yl)cyclohexanol (dihydrocarveol)-   5-methyl-2-(prop-1-en-2-yl)cyclohexanol (isopulegol)    Class of Substituted Cyclohexyl propyl-1,3-diols:-   2-(trans-4-ethylcyclohexyl)propane-1,3-diol    2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol

Class of Menthadiene Alcohols:

-   2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol (Carveol)

Class of Isopropyl Methyl Phenols:

-   5-isopropyl-2-methylphenol-   3-isopropyl-5-methylphenol-   4-isopropyl-3-methylphenol

Class of Monosubstituted Phenols:

-   (E)-2-(prop-1-enyl)phenol,-   4-propylphenol,-   2-tert-butylphenol,-   2-sec-butylphenol,-   2-n-propylphenol,-   3-n-propylphenol,-   4-n-butylphenol,-   4-pentylphenol-   4-sec-butylphenol,-   3-tert-butylphenol.

Monosubstituted phenols generally have the following structure

-   -   wherein        -   the substituent R₁ is selected from the group consisting of            -   linear C₃ to C₅ alkyl,            -   isopropyl            -   branched C₄ alkyl,            -   linear C₃ to C₅ alkenyl,            -   linear C₄ or C₅ alkadienyl,            -   branched C₄ alkenyl,            -   cyclopentyl,            -   cyclopentenyl,            -   cyclohexyl,            -   cyclohexenyl,            -   phenyl, and            -   benzyl.

Isopropyl-methyl-phenols generally have the following structure

Among the phenolic compounds 2-n-propylphenol, 4-n-butylphenol and4-sec-butylphenol are especially preferred since they are evaluated bythe present inventors to be more safe for use in consumer products.Among the antimicrobial alcohols 2-methyl-5-(prop-1-en-2-yl)cyclohexanol(dihydrocarveol), 5-methyl-2-(prop-1-en-2-yl)cyclohexanol (isopulegol)and 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol (carveol) are preferredsince they are evaluated by the present inventors to be more safe foruse in consumer products.

In a preferred embodiment the synergistic antimicrobial compositioncomprises: (a) at least one microbicide selected from the groupconsisting of 2-methyl-5-(prop-1-en-2-yl)cyclohexanol, and5-methyl-2-(prop-1-en-2-yl)cyclohexanol, and (b) at least onemicrobicide selected from the group consisting of 2-n-propylphenol,4-n-butylphenol, and 4-sec-butylphenol.

In another preferred embodiment the synergistic antimicrobialcomposition which comprises: (a)2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol and (b) at least onemicrobicide selected from the group consisting of 2-n-propylphenol,4-n-butylphenol, and 4-sec-butylphenol.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 5-isopropyl-2-methylphenol (alsoknown as carvacrol) and 2-methyl-5-(prop-1-en-2-yl)cyclohexanol (alsoknown as dihydrocarveol). Preferably, a weight ratio of5-isopropyl-2-methylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol isabout 1/3.13.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises (E)-2-(prop-1-enyl)phenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol.

Preferably, a weight ratio of (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/0.06 to 1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-propylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol. Preferably, a weight ratio of4-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol is 1/0.08 to1/5

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-tert-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol. Preferably, a weight ratio of2-tert-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol is about1/3.13.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-n-propylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol. Preferably, a weight ratio of2-n-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from1/0.05 to 1/3.13, preferably from 1/0.05 to 1/0.13 or 1/0.17 to 1/3.13,preferably from 1/0.17 to 1/3.13.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-n-propylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol. Preferably, a weight ratio of3-n-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from1/0.08 to 1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-sec-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol. Preferably, a weight ratio of4-sec-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/1to 1/3.13.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-tert-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol. Preferably, a weight ratio of3-tert-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from1/0.38 to 1/3.13.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 5-isopropyl-2-methylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 5-isopropyl-2-methylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.06 to1/2.5, preferably from 1/0.25 to 1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-isopropyl-3-methylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 4-isopropyl-3-methylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.5 to1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises (E)-2-(prop-1-enyl)phenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of (E)-2-(prop-1-enyl)phenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.13 to1/1.25.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-propylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 4-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.5 to1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-tert-butylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol.

Preferably, a weight ratio of 2-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.17 to1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-sec-butylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 2-sec-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.05 to1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-n-propylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol.

Preferably, a weight ratio of 2-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.05 to1/1.25, preferably from 1/0.25 to 1/1.25.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-n-propylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 3-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.06 to1/2.5, preferably from 1/0.06 to 1/0.013 or 1/0.25 to 1/2.5, preferablyfrom 1/0.25 to 1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-n-butylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 4-n-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.13 to1/2.5, preferably from 1/0.38 to 1/2.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-sec-butylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 4-sec-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is about 1/3.13.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-tert-butylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol. Preferably, a weightratio of 3-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.13 to1/0.5, preferably from 1/0.38 to 1/0.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises2-(trans-4-ethylcyclohexyl)propane-1,3-diol and 4-propylphenol.Preferably, a weight ratio of 4-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol is from 1/0.06 to 1/0.4,preferably from 1/0.06 to 1/0.08 or 1/0.25 to 1/0.4.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises2-(trans-4-ethylcyclohexyl)propane-1,3-diol and 2-n-propylphenol.Preferably, a weight ratio of 2-n-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol is from 1/0.13 to 1/3,preferably from 1/0.13 to 1/0.19 or 1/0.33 to 1/3.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises2-(trans-4-ethylcyclohexyl)propane-1,3-diol and 3-n-propylphenol.Preferably, a weight ratio of 3-n-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol is from 1/0.25 to 1/3,preferably from 1/0.33 to 1/3.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol and 3-n-propylphenol.Preferably, a weight ratio of 3-n-propylphenol to2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol is from 1/0.8 to 1/1.7,preferably from 1/1 to 1/1.7.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 5-isopropyl-2-methylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol (also known as carveol).Preferably, a weight ratio of 5-isopropyl-2-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is about 1/4.4.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-isopropyl-5-methylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 3-isopropyl-5-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.38 to 1/4.38.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-isopropyl-3-methylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 4-isopropyl-3-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.25 to 1/5.8.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises (E)-2-(prop-1-enyl)phenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.17 to 1/1.75.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-propylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 4-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from1/0.08 to 1/4.4.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-tert-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 2-tert-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol isfrom 1/0.38 to 1/4.38.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-sec-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 2-sec-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol isfrom 1/0.38 to 1/4.38.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 2-n-propylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 2-n-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol isfrom 1/0.19 to 1/3.3, preferably 1/0.25 to 1/3.5.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-n-propylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 3-n-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol isfrom 1/0.05 to 1/4.38, preferably from 1/0.05 to 1/0.19 or 1/0.25 to1/4.38, preferably 1/0.25 to 1/4.38.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-n-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 4-n-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from1/1 to 1/11.7.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 4-sec-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 4-sec-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol isfrom 1/4.4 to 1/7.

In a preferred embodiment of the invention, the synergisticantimicrobial composition comprises 3-tert-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol. Preferably, a weight ratioof 3-tert-butylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol isfrom 1/0.38 to 1/5.8.

Combinations according to the invention are capable of very fastantimicrobial action. For instance, we found that complete microbialinactivation could be effected with compositions according to thepresent invention, in most cases, after a contact time of only 15seconds.

The microbicides in the composition of this invention may be used “asis” or may first be formulated with a solvent or a solid carrier.Suitable solvents include, for example, water; glycols, such as ethyleneglycol, propylene glycol, diethylene glycol, dipropylene glycolpolyethylene glycol, and polypropylene glycol; glycol ethers; alcohols,such as methanol, ethanol, propanol, phenethyl alcohol andphenoxypropanol; ketones, such as acetone and methyl ethyl ketone;esters, such as ethyl acetate, butyl acetate, triacetyl citrate, andglycerol triacetate; carbonates, such as propylene carbonate anddimethyl carbonate; inorganic particulate material, starch, air andmixtures thereof. In certain preferred embodiments, suitable solventsinclude for example water, glycols, glycol ethers, esters and mixturesthereof. Suitable solid carriers include, for example, cyclodextrin,silicas, clays, talc, calcite, dolomite, aluminosilicate, diatomaceousearth, waxes, cellulosic materials, alkali and alkaline earth (e.g.,sodium, magnesium, potassium) metal salts (e.g., chloride, nitrate,bromide, sulfate) and charcoal.

Particularly preferred carriers are water or oil/solvent and even morepreferred is a carrier that is a mixture of water and oil. Examples ofoils include mineral oils, oils of biological origin (e.g. vegetableoils), and petroleum-derived oils and waxes. The oils of biologicalorigin are preferably triglyceride-based. Preferably, the carrier oil isnot a perfume oil. Thus, the carrier oil preferably does notsubstantially contribute to the odour of the composition, morepreferably it does not contribute to that odour. Examples of solventsinclude alcohols, ethers and acetone. The starch may be natural starchobtained from food grains or may be a modified starch.

Air can for instance be used as a carrier when the components accordingto the invention and/or the terpineol are atomised or otherwisedispersed as a fine mist.

Particularly preferred carriers are water or oil/solvent and even morepreferred is a carrier that is a mixture of water and oil. Thus, in manyof the envisaged applications like personal care/washing, oral care andhard surface cleaning, the antimicrobial composition may be formulatedwith either an aqueous base or an oil/solvent base. Compositions with anaqueous base (water being the carrier), can also for instance beproducts in gel format. Compositions with an oil/solvent base can forinstance be products in anhydrous stick form or propellant-containingproducts.

Thus, the antimicrobial composition can for instance, preferably be anantimicrobial anhydrous stick personal care composition on anoil/solvent base wherein the composition has a water content of lessthan 0.01% by weight, and wherein the composition preferably is free ofwater. Alternatively, the antimicrobial composition can for instance,preferably be an antimicrobial propellant-drivable personal carecomposition, also comprising a propellant. Air can also be used aspropellant, for instance in the form of compressed or liquefied air.

However, the most preferred product format has an emulsion base (waterand/or oil being the carrier) or is capable of forming an emulsion upondilution, e.g. soap products in liquid, solid, lotion or semisolid formfor hand wash, face wash, body wash, or shaving applications;toothpaste/dentifrices for oral care applications or products for hardsurface cleaning in bars or liquids form. If the product comprises anemulsion base, it preferably also comprises one or more surfactants asdescribed below.

“Substantially free” means, e.g., having less than 5 wt % based on theweight of active ingredients (i.e., the weight of claimed components a)and b) plus the additional ingredients listed in this paragraph),preferably less than 3 wt %, preferably less than 1 wt %, preferablyless than 0.5 wt %, preferably less than 0.2 wt %.

When a microbicide component is formulated in a solvent, the formulationmay optionally contain surfactants. When such formulations containsurfactants, they can be in the form of emulsive concentrates,emulsions, microemulsive concentrates, or microemulsions. Emulsiveconcentrates form emulsions upon the addition of a sufficient amount ofwater. Microemulsive concentrates form microemulsions upon the additionof a sufficient amount of water. Such emulsive and microemulsiveconcentrates are generally well known in the art. U.S. Pat. No.5,444,078 may be consulted for further general and specific details onthe preparation of various microemulsions and microemulsiveconcentrates.

A preferred product format has an emulsion base (water and/or oil beingthe carrier) or is capable of forming an emulsion upon dilution, e.g.soap products in liquid, solid, lotion or semisolid form for hand wash,face wash, body wash, or shaving applications; toothpaste/dentifricesfor oral care applications or products for hard surface cleaning in barsor liquids form. If the product comprises an emulsion base, itpreferably also comprises one or more surfactants as described below.

It is particularly preferred that the microbiocidal compositioncomprises from 1 to 80% by weight of one or more surfactants in additionto the synergistic combination of microbiocides claimed in the presentinvention.

In general, the surfactants may be chosen from the surfactants describedin well-known textbooks like “Surface Active Agents” Vol. 1, by Schwartz& Perry, Interscience 1949, Vol. 2 by Schwartz, Perry & Berch,Interscience 1958, and/or the current edition of “McCutcheon'sEmulsifiers and Detergents” published by Manufacturing ConfectionersCompany or in “Tenside-Taschenbuch”, H. Stache, 2nd Edn., Carl HauserVerlag, 1981; “Handbook of Industrial Surfactants” (4th Edn.) by MichaelAsh and Irene Ash; Synapse Information Resources, 2008. Any type ofsurfactant, i.e. anionic, cationic, nonionic, zwitterionic or amphotericcan be used. Preferably, the one or more surfactants are anionic,nonionic, or a combination of anionic and nonionic surfactants. Morepreferably, the one or more surfactants are anionic.

A particularly preferred surfactant is soap. Soap is a suitablesurfactant for personal washing applications of the antimicrobialcomposition of the invention. The soap is preferably C₈-C₂₄ soap, morepreferably a C₁₀-C₂₀ soap and most preferably C₁₂-C₁₆ soap. The soap mayor may not have one or more carbon-carbon double bonds or triple bonds.The cation of the soap can for instance be an alkali metal, alkalineearth metal or ammonium. Preferably, the cation of the soap is selectedfrom sodium, potassium or ammonium. More preferably the cation of thesoap is sodium or potassium.

The soap may be obtained by saponifying a fat and/or a fatty acid. Thefats or oils may be fats or oils generally used in soap manufacture,such as tallow, tallow stearines, palm oil, palm stearines, soya beanoil, fish oil, castor oil, rice bran oil, sunflower oil, coconut oil,babassu oil, palm kernel oil, and others. In the above process the fattyacids are derived from oils/fats selected from coconut, rice bran,groundnut, tallow, palm, palm kernel, cotton seed, soyabean, castor etc.The fatty acid soaps can also be synthetically prepared (e.g. by theoxidation of petroleum or by the hydrogenation of carbon monoxide by theFischer-Tropsch process). Resin acids, such as those present in talloil, may be used. Naphthenic acids are also suitable.

Tallow fatty acids can be derived from various animal sources. Othersimilar mixtures, such as those from palm oil and those derived fromvarious animal tallow and lard are also included.

A typical fatty acid blend consists of 5 to 30%-wt coconut fatty acidsand 70 to 95%-wt fatty acids ex hardened rice bran oil. Fatty acidsderived from other suitable oils/fats such as groundnut, soybean,tallow, palm, palm kernel, etc. may also be used in other desiredproportions. The soap, when present in solid forms of the presentinvention, is preferably present in an amount of 30 to 80%, morepreferably from 50 to 80%, and even more preferably 55 to 75% by weightof the composition. The soap, when present in liquid forms of thecomposition is preferably present in 0.5 to 20%., more preferably from 1to 10% by weight of the composition.

Other preferred surfactants fatty acid glycinates and fattyamphocarboxylates. These surfactants are particularly preferred in skinand hair cleaning compositions, because of their mild detergency andhighly foaming nature. The fatty acid glycinates are salts of fatty acidamides of glycine, including for example sodium cocoyl glycinate. Thefatty amphocarboxylates are amphoteric surfactants including for examplesodium lauroamphoacetate (i.e. sodium2-[1-(2-hydroxyethyl)-2-undecyl-4,5-dihydroimidazol-1-ium-1-yl]acetate).Yet another example of suitable surfactants are derivatives ofisethionates, including acylisethionates.

The antimicrobial composition of the invention is also useful in hardsurface cleaning applications. In such applications, preferredsurfactants are nonionic surfactants, such as C₈-C₂₂, preferably C₈-C₁₆fatty alcohol ethoxylates, comprising between 1 and 8 ethylene oxidegroups when the product is in the liquid form. When the product for hardsurface cleaning applications is in the solid form, surfactants arepreferably selected from primary alkyl sulphates, secondary alkylsulphonates, alkyl benzene sulphonates, ethoxylated alkyl sulphates, oralcohol ethoxylate nonionic surfactants. The composition may furthercomprise an anionic surfactant, such as alkyl ether sulphate preferablythose having between 1 and 3 ethylene oxide groups, either from naturalor synthetic source and/or sulphonic acid. Especially preferred aresodium lauryl ether sulphates. Alkyl polyglucoside may also be presentin the composition, preferably those having a carbon chain lengthbetween C6 and C16. Other classes of useful surfactants include cationicsurfactants, such as long chain quaternary ammonium compounds andamphoteric surfactants such as betaines and alkyl dimethyl amine oxides.Suitable surfactant concentrations in liquid forms of hard surfacecleaning application are generally from about from 0.5 to 10%,preferably from 1 to 5% by weight of the composition. In solidcompositions, surfactant is preferably present in 5 to 40%, preferablyfrom 10 to 30% by weight of the composition.

The antimicrobial composition of the invention is useful in oral carecompositions e.g. in a dentifrice/toothpaste or an oral rinse product.In such applications, preferred surfactants are anionic, nonionic oramphoteric in nature, preferably anionic or amphoteric. The anionicsurfactant is preferably an alkali metal alkyl sulphate, more preferablya sodium lauryl sulphate (SLS). Mixtures of anionic surfactants may alsobe employed. The amphoteric surfactant is preferably a betaine, morepreferably an alkylamidopropyl betaine (wherein the alkyl group is alinear C₁₀-C₁₈ chain), and most preferably is cocoamidopropyl betaine(CAPB). Mixtures of amphoteric surfactants may also be employed.Suitable surfactant concentrations in oral care application aregenerally from about 2% to about 15%, preferably from about 2.2% toabout 10%, more preferably from about 2.5 to about 5% by weight of thetotal composition.

Thus, it is highly preferred that the antimicrobial compositions includesoap, alkyl sulphate or linear alkyl benzene sulphonate as thesurfactants. More preferably, the one or more surfactants are selectedfrom the group consisting of soaps, alkyl sulphates and linear alkylbenzene sulphonates.

A microbicide component also can be formulated in the form of adispersion. The solvent component of the dispersion can be an organicsolvent or water, preferably water. Such dispersions can containadjuvants, for example, co-solvents, thickeners, anti-freeze agents,dispersants, fillers, pigments, surfactants, biodispersants,polycations, stabilizers, scale inhibitors and anti-corrosion additives.

When both microbicides are each first formulated with a solvent, thesolvent used for the first microbicide may be the same as or differentfrom the solvent used to formulate the other commercial microbicide,although water is preferred for most industrial biocide applications. Itis preferred that the two solvents are miscible.

The composition may further comprise various additional ingredientsknown to a person skilled in the art. Such additional ingredientsinclude but are not limited to: perfumes, pigments, preservative,emollients, sunscreens, emulsifiers, gelling agents, thickening agents,humectants (e.g. glycerine, sorbitol), sequestrants (e.g. EDTA) orpolymers (e.g. cellulose derivatives for structuring such as methylcellulose)

The antimicrobial composition may be in form of a solid, a liquid, a gelor a paste. A person skilled in the art can prepare compositions invarious formats by choosing one or more carrier materials and/orsurfactant. The antimicrobial compositions of the present invention areuseful for cleansing and care, in particular for skin cleansing and skincare. It is envisaged that the antimicrobial composition can be used asa leave-on product or a wash-off product, preferably a wash-off product.The antimicrobial composition of the present invention can also be usedfor cleansing and care of hard surfaces such as glass, metal, plasticand the like.

Those skilled in the art will recognize that the microbicide componentsof the present invention may be added to a locus sequentially,simultaneously, or may be combined before being added to the locus. Itis preferred that the first microbicide and the second microbicidecomponent be added to a locus simultaneously or sequentially. When themicrobicides are added simultaneously or sequentially each individualcomponent may contain adjuvants, such as, for example, solvent,thickeners, anti-freeze agents, colorants, sequestrants (such asethylenediamine-tetraacetic acid, ethylenediaminedisuccinic acid,iminodisuccinic acid and salts thereof), dispersants, surfactants,biodispersants, polycations, stabilizers, scale inhibitors andanti-corrosion additives.

The microbicide compositions of the present invention can be used toinhibit the growth of or kill microorganisms by introducing amicrobicidally effective amount of the compositions onto, into or at alocus subject to attack.

Suitable loci include, for example: industrial process water includingelectrocoat deposition systems, cooling towers and air washers; gasscrubbers; wastewater treatment; ornamental fountains; reverse osmosisfiltration; ultrafiltration; ballast water; evaporative condensers andheat exchangers; pulp and paper processing fluids and additives; mineralslurries; starch; plastics; emulsions; dispersions; paints; latices;coatings, such as varnishes; construction products, such as mastics,caulks, and sealants; construction adhesives, such as ceramic adhesives,carpet backing adhesives, and laminating adhesives; industrial orconsumer adhesives; photographic chemicals; printing fluids; householdand institutional products used in restaurants, healthcare facilities,schools, food processing facilities and farms including, cleaners,sanitizers and disinfectants, wipes, soaps, detergents, floor polishesand laundry rinse water; cosmetics; toiletries; shampoos; metalworkingfluids; conveyor lubricants; hydraulic fluids; leather and leatherprocessing products; textiles; textile and textile processing products;wood and wood processing products, such as plywood, chipboard,wallboard, flakeboard, laminated beams, oriented strandboard, hardboard,and particleboard; oil and gas processing fluids such as injectionfluids, fracture fluids, drilling muds and produced water; fueltransportation and storage systems; agriculture adjuvant preservation;surfactant preservation; medical devices; diagnostic reagentpreservation; food preservation, such as plastic or paper food wrap;food, beverage, and industrial process pasteurizers; toilet bowls;recreational water; pools; and spas.

In preferred embodiments, the composition is particularly suited forapplication to the skin. For example, a surface like the hands, face,body, or the oral cavity can suitably be contacted with the compositionof the invention. In other preferred embodiments, the surface is anyhard surface. Typically, such hard surfaces are surfaces that commonlyrequire cleaning and often also require sanitization or disinfection.Such surfaces can be found in many household or industrial environments,and can include for example kitchen and bathroom surfaces, table tops,floors, walls, windows, utensils, cutlery, and crockery. Such surfacescan be made from many different materials, for instance plastics, wood,metal, ceramics, glass, concrete, marble, and painted surfaces. Inpreferred embodiments, the compositions can be used for suchdisinfection, reduction in microbial count or improved hygiene at lociother than the surfaces as described hereinbefore.

In preferred embodiments, the invention relates to compositionsaccording to the invention for use as or incorporation in home careproducts and personal care products. More preferably, this embodiment ofthe invention relates to a composition according to the invention whichis a home care product or a personal care product.

A “home care product” is a product used for the treatment, cleaning,caring or conditioning of the home or any of its contents. The foregoingincludes, but is not limited to, compositions, products, or combinationsthereof relating to or having use or application in the treatment,cleaning, cleansing, caring or conditioning of surfaces, furniture andatmosphere of the home and household contents, such as clothes, fabricsand/or cloth fibers and the manufacture of all of the foregoingproducts. A “personal care product” is a product for the treatment,cleaning, caring or conditioning of the person. The foregoing includes,but is not limited to, chemicals, compositions, products, orcombinations thereof relating to or having use or application in thetreatment, cleaning, cleansing or conditioning of the person (includingin particular the skin, hair and oral cavity), and the manufacture ofall the foregoing. Home care products and personal care products are forexample products marketed under mass market brands, non-limitingexamples being soap bars, deodorants, shampoos, and home surfacesanitizers/disinfectants.

According to another aspect of the invention, there is provided a methodof disinfecting a surface comprising the steps of

-   -   a. applying a composition according to the invention on to the        surface; and    -   b. removing the composition from the surface.

The method according to the present invention also includes the step ofremoving the composition from the surface. Here, removing thecomposition also encompasses partially removing the composition, becausetraces of the composition may remain on the surface. In many typicalsituations, such as washing of the skin or hard-surface cleaning, it isacceptable or sometimes even desirable if part of the composition—inparticular certain active ingredients—remains on the surface. Therefore,step b preferably involves removing at least 5%, more preferably atleast 10%, even more preferably at least 25%, still more preferably atleast 50% and yet more preferably at least 75% of the composition byweight. Preferably, the step of removing the composition comprisesrinsing the surface with a suitable solvent or wiping the surface with asuitable wipe, more preferably, this step consists of rinsing thesurface with a suitable solvent or wiping the surface with a suitablewipe. Alternatively, the removal step can also include evaporation ofpart of the composition, for example when the composition comprisesvolatile components. e.g. solvents.

If the surface is a surface of a human or animal body, the methodpreferably is a non-therapeutic method of disinfecting a surface.

A suitable medium for rinsing the surface is water but it could also befor example a mixture of water and alcohol. It is then rinsed preferablywith sufficient amounts of water after a pre-determined period of timeto remove any visible or sensory residue of the composition.Alternatively, an alcohol wipe or a water/alcohol impregnated wipe maybe used to wipe the surface to be visibly free of the anti-microbialcomposition. The step of removing the composition (e.g. by rinsing orwiping the surface) is preferably started less than 5 minutes, morepreferably less than 2 minutes, even more preferably less than 1 minute,still more preferably less than 30 seconds and yet more preferably lessthan 20 seconds after commencement of the step of applying thecomposition on the surface, because of the surprisingly fastantimicrobial action of the compositions according to the presentinvention. Even though partial microbial kill may be almostinstantaneous upon application of the composition according to theinvention, it is preferred that the step of removing the compositionfrom the surface is started out at least 5 seconds, preferably at least10 seconds, more preferably at least 15 seconds after commencement ofthe step of applying the composition on the surface, in order to effectoptimal antimicrobial action. Combinations of these times into timeintervals are preferred too. Therefore, it is particularly preferredthat the step of removing the composition from the surface (i.e. step b)is started between 2 minutes and 5 seconds, more preferably between 1minute and 10 seconds, even more preferably between 30 and 10 secondsand still more preferably between 20 and 15 seconds after commencementof the step of applying the composition on the surface (i.e. step a).

Disinfection Time

These times between applying the composition and rinsing or wiping arepreferably related to the disinfection time, in order to ensure optimalcleansing and sanitising of the surface. Therefore, the inventionpreferably relates to a method, wherein the disinfection time T of saidmethod is less than 300 seconds, preferably less than 60 seconds, andmore preferably less than 15 seconds; wherein T is defined as the timethat elapses from the moment of adding the composition to a microbialculture until the number of microbes per unit volume of the culture isreduced by a factor of 100 000; and wherein the initial number ofmicrobes preferably exceeds about 100 000 000 microbes per millilitreand wherein the composition is preferably a liquid composition.

The disinfecting action of the method (as may be expressed in terms ofthe disinfection time T) is preferably determined according to theprotocol of Example 1 as described hereinafter. This test relates to astandardised test environment in which the microbial culture is kept insuspension. A similarly suitable test is the standard suspension methoddescribed in European Standard EN1276, with the proviso that thedisinfection time is adapted to suit the above criteria as will be clearto a person skilled in the art. Alternatively, one of the test methodsas described in WO 2010/046238 may for instance be applied to establishthe disinfecting action.

Such test methods may preferably also be used by the skilled person todetermine the optimal concentrations of the one or more components in anantimicrobial composition according to the present invention.

Alternatively, since the method is directed towards surfacedisinfection, the disinfection time may also be determined by testmethods involving a surface. Therefore, the invention preferably relatesto a method according to the present invention, wherein the surfacedisinfection time T2 of said method is less than 60 seconds, preferablyless than 15 seconds, wherein T2 is defined as the time starting fromthe moment of applying the composition to the surface to be disinfectedafter which the number of microbes per unit area is reduced by a factorof 10000 (i.e. a 4 log reduction), wherein the initial number ofmicrobes preferably exceeds 10³, more preferably 10⁵, and even morepreferably 10⁷ microbes per square centimetre. Such tests may forinstance be performed as described in WO 2010/046238, or as described inEuropean Standards EN 13697:2001 and EN 1500:1997.

Another preferred embodiment of the invention relates to compositionsaccording to the invention for use as or incorporation in industrialand/or institutional products. More preferably, this embodiment of theinvention relates to a composition according to the invention which isan industrial and/or an institutional product. Industrial andinstitutional products are for example products being marketed underprofessional brands, non-limiting examples being for industrial,institutional, janitorial, and medical cleaning, cleaning-in-place, foodservices, veterinary, and agricultural products. Industrial and/orinstitutional products also include products for cleaning of the person(such as hand sanitizers) for medical offices, hospitals and/or otherinstitutions.

In another preferred embodiment, the invention also relates to a methodor use according to the invention involving home care products orpersonal care products. For example, the method according to theinvention—which comprises application of a composition according to theinvention in step a—can be a method wherein that composition is acomposition for use as or incorporation in home care products andpersonal care products as described hereinabove. Similarly, in anotherpreferred embodiment, the invention also relates to a method or useaccording to the invention involving industrial and/or institutionalproducts. For example, the method according to the invention—whichcomprises application of a composition according to the invention instep a—can be a method wherein that composition is a composition for useas or incorporation in industrial and/or institutional products asdescribed hereinabove.

Products and/or methods for use in the home care or personal care fieldare generally distinct from products and/or methods for use in theindustrial and/or institutional field. Thus, for example, a product thatis marketed as a home or personal care product will generally not bemarketed as a product for industrial and/or institutional use and viceversa. Therefore, certain preferred embodiments of the presentinvention, when carried forth into practice, will relate to the onefield, but not the other.

The specific amount of the composition of this invention necessary toinhibit or control the growth of microorganisms in a locus depends uponthe particular locus to be protected. Typically, the amount of thecomposition of the present invention to control the growth ofmicroorganisms in a locus is sufficient if it provides a total of from0.02 to 4% of the microbicide ingredients of the composition in thelocus. It is preferred that the microbicide ingredients of thecomposition be present in the locus in an amount of at least 0.05%,preferably at least 0.1%, preferably at least 0.2%, preferably at least0.3%, preferably at least 0.4%. It is preferred that the microbicideingredients of the composition be present in the locus at a total amountof no more than 4%, preferably no more than 2%, preferably no more than1%.

EXAMPLES Materials and Methods

The synergy of the combinations of the present invention wasdemonstrated by testing a wide range of concentrations and ratios of thecompounds against the noted organism. One skilled in the art willrecognize that the sensitivity of other microorganisms to the particularcombinations will vary and, as a result, the concentrations, the ratios,for each, or both, of the compounds may vary from those detailed inthese examples. The concentrations and ratios may also vary underdifferent test conditions or with different test methods.

One measure of synergy is the industrially accepted method described byKull, F. C.; Eisman. P. C.; Sylwestrowicz, H. D. and Mayer, R. L., inApplied Microbiology 9:538-541 (1961), using the ratio determined by theformula:

Q _(a) /Q _(A) ÷Q _(b) /Q _(B)=Synergy Index (“SI”)

wherein:Q_(A)=concentration of compound A (first component) in percent, actingalone, which produced an end point (MBC of Compound A).Q_(B)=concentration of compound A in percent, in the mixture, whichproduced an end point.Q_(B)=concentration of compound B (second component) in percent, actingalone, which produced an end point (MBC of Compound B).Q_(b)=concentration of compound B in percent, in the mixture, whichproduced an end point.

When the sum of Q_(a)/Q_(A) and Q_(b)/Q_(B) is greater than one,antagonism is indicated. When the sum is equal to one, additivity isindicated, and when less than one, synergy is demonstrated. The lowerthe SI, the greater is the synergy shown by that particular mixture. Theminimum biocidal concentration (MBC) of a microbicide is the lowestconcentration tested under a specific set of conditions that providescomplete kill of the tested microorganisms.

Synergy tests were conducted using standard microtiter plate assays withphosphate buffer containing 35% dipropylene glycol (DPG). In thismethod, a wide range of combinations of chemicals was tested byconducting high resolution MBC assays of Component (A) in the presenceof various concentrations of Component (B). High resolution MBCs weredetermined by adding varying amounts of microbicide to one column of amicrotiter plate and doing subsequent ten-fold dilutions using anautomated liquid handling system to obtain a series of closely spacedendpoints. The MBC plate was inoculated one column at a time with thetest microorganism. An aliquot of the inoculated well was transferred at15 seconds to a plate containing a neutralizing agent (D/E NeutralizingBroth), mixed and held for 5 minutes before being transferred to agrowth plate containing trypticase soy broth (TSB). The TSB plate wasincubated at 37° C. and read for the presence/absence of growth at 24hours. The lowest level tested that provided complete kill (as evidencedby no growth in the microtitre plate) of the test organisms in 15seconds is the minimum biocidal concentration (MBC).

The synergy of the combinations of the present invention was determinedagainst a bacterium, Esherichia coli (E. coli—ATCC #10536), at aconcentration of approximately 1×10⁸ bacteria per mL. This microorganismis representative of natural contaminants in many consumer andindustrial applications. The plates were visually evaluated formicrobial growth (turbidity) to determine the MBC after 24 hoursincubation time at 37° C.

The test results for demonstration of synergy of the combinations of thepresent invention are shown below in Tables 1 through 38. Each tableshows the specific combinations of the two components; results againstthe microorganism tested; the end-point activity in weight % measured bythe MBC for the first component alone (Q_(A)), for the second componentalone (Q_(B)), for the first component in the mixture (Q_(a)) and forthe second component in the mixture (Q_(b)); the calculated SI value;and the range of synergistic ratios for each combination tested (firstcomponent to second component or A/B) against the particularmicroorganism.

Data in the tables below include the range of ratios that were found tobe synergistic. (Data which were collected for combinations whereconcentrations were equal to or greater than Q_(A) or Q_(B) are notreported.) These data demonstrate that certain combinations ofcomponents A and B show enhanced control over the microorganisms thanwould be expected if the combinations were additive rather thansynergistic.

TABLE 1 First Component (A) = 5-isopropyl-2-methylphenol SecondComponent (B) = 2-methyl-5-(prop-1-en-2-yl)cyclohexanol MicroorganismQ_(a) Q_(b) SI Ratio A to B E. coli 10536 0.2 0 1.00 0.08 0.25 0.90 1 to3.13 0 0.5 1.00The ratios of 5-isopropyl-2-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratio of 5-isopropyl-2-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is 1/3.13.

TABLE 2 First Component (A) = (E)-2-(prop-1-enyl)phenol Second Component(B) = 2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b)SI Ratio A to B E. coli 10536 0.5 0 1.00 0.3 0.025 0.65 1 to 0.08 0.40.025 0.85 1 to 0.06 0.2 0.05 0.50 1 to 0.25 0.3 0.05 0.70 1 to 0.17 0.40.05 0.90 1 to 0.13 0.2 0.075 0.55 1 to 0.38 0.3 0.075 0.75 1 to 0.250.4 0.075 0.95 1 to 0.19 0.2 0.1 0.60 1 to 0.50 0.3 0.1 0.80 1 to 0.330.1 0.25 0.70 1 to 2.50 0.2 0.25 0.90 1 to 1.25 0 0.5 1.00The ratios of (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratios of (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol range from 1/0.06 to 1/2.5.

TABLE 3 First Component (A) = 4-propylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.4 0 1.00 0.2 0.025 0.55 1 to 0.13 0.3 0.0250.80 1 to 0.08 0.2 0.05 0.60 1 to 0.25 0.3 0.05 0.85 1 to 0.17 0.2 0.0750.65 1 to 0.38 0.3 0.075 0.90 1 to 0.25 0.2 0.1 0.70 1 to 0.50 0.3 0.10.95 1 to 0.33 0.05 0.25 0.63 1 to 5 0.06 0.25 0.65 1 to 4.17 0.08 0.250.70 1 to 3.13 0.1 0.25 0.75 1 to 2.50 0 0.5 1.00The ratios of 4-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanoltested ranged from 1/0.025 to 1/350. The synergistic ratios of4-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol range from1/0.08 to 1/5.

TABLE 4 First Component (A) = 2-tert-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.2 0 1.00 0.08 0.25 0.90 1/3.13 0 0.5 1.00The ratios of 2-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratio of 2-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is 1/3.13.

TABLE 5 First Component (A) = 2-n-propylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.6 0 1.00 0.2 0.025 0.38 1 to 0.13 0.3 0.0250.55 1 to 0.08 0.4 0.025 0.72 1 to 0.06 0.5 0.025 0.88 1 to 0.05 0.20.05 0.43 1 to 0.25 0.3 0.05 0.60 1 to 0.17 0.4 0.05 0.77 1 to 0.13 0.50.05 0.93 1 to 0.10 0.2 0.075 0.48 1 to 0.38 0.3 0.075 0.65 1 to 0.250.4 0.075 0.82 1 to 0.19 0.5 0.075 0.98 1 to 0.15 0.2 0.1 0.53 1 to 0.500.3 0.1 0.70 1 to 0.33 0.4 0.1 0.87 1 to 0.25 0.06 0.1 0.30 1 to 1.670.08 0.25 0.63 1 to 3.13 0.1 0.25 0.67 1 to 2.50 0.2 0.25 0.83 1 to 1.250 0.5 1.00The ratios of 2-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratios of 2-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol range from 1/0.05 to 1/3.13.

TABLE 6 First Component (A) = 3-n-propylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.4 0 1.00 0.3 0.025 0.80 1 to 0.08 0.3 0.050.85 1 to 0.17 0.3 0.075 0.90 1 to 0.25 0.2 0.1 0.70 1 to 0.50 0.1 0.250.75 1 to 2.50 0 0.5 1.00The ratios of 3-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratios of 3-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol range from 1/0.08 to 1/2.5.

TABLE 7 First Component (A) = 4-sec-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.2 0 1.00 0.1 0.1 0.70 1 to 1 0.08 0.25 0.901 to 3.13 0 0.5 1.00The ratios of 4-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratios of 4-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol range from 1/1 to 1/3.13.

TABLE 8 First Component (A) = 3-tert-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohexanol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.3 0 1.00 0.2 0.075 0.82 1 to 0.38 0.2 0.10.87 1 to 0.50 0.08 0.25 0.77 1 to 3.13 0.1 0.25 0.83 1 to 2.50 0 0.51.00The ratios of 3-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ranged from 1/0.025 to1/350. The synergistic ratios of 3-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol range from 1/0.38 to 1/3.13.

TABLE 9 First Component (A) = 5-isopropyl-2-methylphenol SecondComponent (B) = (1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanolMicroorganism Q_(a) Q_(b) SI Ratio A to B E. coli 10536 0.5 0 1.00 0.30.025 0.65 1 to 0.08 0.4 0.025 0.85 1 to 0.06 0.3 0.05 0.70 1 to 0.170.4 0.05 0.90 1 to 0.13 0.2 0.075 0.55 1 to 0.38 0.3 0.075 0.75 1 to0.25 0.4 0.075 0.95 1 to 0.19 0.2 0.1 0.60 1 to 0.50 0.3 0.1 0.80 1 to0.33 0.1 0.25 0.70 1 to 2.50 0.2 0.25 0.90 1 to 1.25 0 0.5 1.00The ratios of 5-isopropyl-2-methylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 5-isopropyl-2-methylphenolto (1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.06to 1/2.5.

TABLE 10 First Component (A) = 4-isopropyl-3-methylphenol SecondComponent (B) = (1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanolMicroorganism Q_(a) Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.20.1 0.87 1 to 0.50 0.1 0.25 0.83 1 to 2.50 0 0.5 1.00The ratios of 4-isopropyl-3-methylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 4-isopropyl-3-methylphenolto (1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.5to 1/2.5.

TABLE 11 First Component (A) = (E)-2-(prop-1-enyl)phenol SecondComponent (B) = (1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanolMicroorganism Q_(a) Q_(b) SI Ratio A to B E. coli 10536 0.8 0 1.00 0.60.075 0.90 1 to 0.13 0.6 0.1 0.95 1 to 0.17 0.2 0.25 0.75 1 to 1.25 0.30.25 0.88 1 to 0.83 0 0.5 1.00The ratios of (E)-2-(prop-1-enyl)phenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of (E)-2-(prop-1-enyl)phenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.13 to1/1.25.

TABLE 12 First Component (A) = 4-propylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.1 0.87 1 to 0.500.1 0.25 0.83 1 to 2.50 0 0.5 1.00The ratios of 4-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 4-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.5 to1/2.5.

TABLE 13 First Component (A) = 2-tert-butylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.4 0 1.00 0.2 0.05 0.60 1 to 0.250.3 0.05 0.85 1 to 0.17 0.3 0.075 0.90 1 to 0.25 0.2 0.1 0.70 1 to 0.500.3 0.1 0.95 1 to 0.33 0.1 0.25 0.75 1 to 2.5 0 0.5 1.00The ratios of 2-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 2-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.17 to1/2.5.

TABLE 14 First Component (A) = 2-sec-butylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.1 0.87 1 to 0.500.1 0.25 0.83 1 to 2.5 0 0.5 1.00The ratios of 2-sec-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 2-sec-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.5 to1/2.5.

TABLE 15 First Component (A) = 2-n-propylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.6 0 1.00 0.3 0.025 0.55 1 to 0.080.4 0.025 0.72 1 to 0.06 0.5 0.025 0.88 1 to 0.05 0.2 0.1 0.53 1 to 0.500.3 0.1 0.70 1 to 0.33 0.4 0.1 0.87 1 to 0.25 0.2 0.25 0.83 1 to 1.25 00.5 1.00The ratios of 2-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 2-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.05 to1/1.25.

TABLE 16 First Component (A) = 3-n-propylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.5 0 1.00 0.3 0.025 0.65 1 to 0.080.4 0.025 0.85 1 to 0.06 0.4 0.05 0.90 1 to 0.13 0.2 0.075 0.55 1 to0.38 0.3 0.075 0.75 1 to 0.25 0.4 0.075 0.95 1 to 0.19 0.2 0.1 0.60 1 to0.50 0.3 0.1 0.80 1 to 0.33 0.1 0.25 0.70 1 to 2.50 0.2 0.25 0.90 1 to1.25 0 0.5 1.00The ratios of 3-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 3-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.06 to1:2.5.

TABLE 17 First Component (A) = 4-n-butylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.025 0.72 1 to 0.130.2 0.075 0.82 1 to 0.38 0.1 0.1 0.53 1 to 1 0.2 0.1 0.87 1 to 0.50 0.10.25 0.83 1 to 2.50 0 0.5 1.00The ratios of 4-n-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 4-n-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 110.13 to1/2.5

TABLE 18 First Component (A) = 4-sec-butylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.2 0 1.00 0.08 0.25 0.90 1 to 3.130 0.5 1.00The ratios of 4-sec butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged fromV/0.025 to 1/350. The synergistic ratio of 4-sec-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is 1/3.13.

TABLE 19 First Component (A) = 3-tert-butylphenol Second Component (B) =(1R,2S,5R)-5-methyl-2-(prop-1- en-2-yl)cyclohexanol Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.025 0.72 1 to 0.130.2 0.075 0.82 1 to 0.38 0.2 0.1 0.87 1 to 0.50 0.2 0.25 1.17 1 to 1.250 0.5 1.00The ratios of 3-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested ranged from1/0.025 to 1/350. The synergistic ratios of 3-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol range from 1/0.13 to1/0.5.

TABLE 20 First Component (A) = 5-isopropyl-2-methylphenol SecondComponent (B) = 2-methyl-5-(prop-1- en-2-yl)cyclohex-2-enolMicroorganism Q_(a) Q_(b) SI Ratio A to B E. coli 10536 0.2 0 1.00 0.080.35 0.90 1 to 4.4 0 0.7 1.00The ratios of 5-isopropyl-2-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratio of 5-isopropyl-2-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is 1/4.4.

TABLE 21 First Component (A) = 3-isopropyl-5-methylphenol SecondComponent (B) = 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol MicroorganismQ_(a) Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.075 0.77 1 to0.38 0.2 0.1 0.81 1 to 0.5 0.08 0.35 0.77 1 to 4.38 0.1 0.35 0.83 1 to3.5 0 0.7 1.00The ratios of 3-isopropyl-5-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 3-isopropyl-5-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.38 to 1/4.4.

TABLE 22 First Component (A) = 4-isopropyl-3-methylphenol SecondComponent (B) = 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol MicroorganismQ_(a) Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.05 0.74 1 to0.25 0.2 0.075 0.77 1 to 0.38 0.2 0.1 0.81 1 to 0.5 0.06 0.35 0.70 1 to5.8 0.08 0.35 0.77 1 to 4.4 0.1 0.35 0.83 1 to 3.5 0 0.7 1.00The ratios of 4-isopropyl-3-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 4-isopropyl-3-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.25 to 1/5.8.

TABLE 23 First Component (A) = (E)-2-(prop-1-enyl)phenol SecondComponent (B) = 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol MicroorganismQ_(a) Q_(b) SI Ratio A to B E. coli 10536 0.8 0 1.00 0.6 0.1 0.89 1 to0.17 0.2 0.35 0.75 1 to 1.75 0.3 0.35 0.88 1 to 1.17 0 0.7 1.00The ratios of (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of (E-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.17 to 1/1.75.

TABLE 24 First Component (A) = 4-propylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.4 0 1.00 0.3 0.025 0.79 1 to 0.08 0.3 0.050.82 1 to 0.17 0.3 0.075 0.86 1 to 0.25 0.2 0.1 0.64 1 to 0.5 0.3 0.10.89 1 to 0.33 0.08 0.35 0.70 1 to 4.4 0.1 0.35 0.75 1 to 3.5 0 0.7 1.00The ratios of 4-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 4-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.08 to 1/4.4.

TABLE 25 First Component (A) = 2-tert-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.3 0 1.00 0.2 0.075 0.77 1 to 0.38 0.2 0.10.81 1 to 0.5 0.08 0.35 0.77 1 to 4.38 0.1 0.35 0.83 1 to 3.5 0 0.7 1.00The ratios of 2-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 2-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.38 to 1/4.4.

TABLE 26 First Component (A) = 2-sec-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.3 0 1.00 0.2 0.075 0.77 1 to 0.38 0.2 0.10.81 1 to 0.5 0.08 0.35 0.77 1 to 4.38 0.1 0.35 0.83 1 to 3.5 0 0.7 1.00The ratios of 2-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 2-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.38 to 1/4.4.

TABLE 27 First Component (A) = 2-n-propylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.6 0 1.00 0.3 0.075 0.61 1 to 0.25 0.4 0.0750.77 1 to 0.19 0.3 0.1 0.64 1 to 0.33 0.5 0.1 0.98 1 to 0.2 0.1 0.350.67 1 to 3.5 0.2 0.35 0.83 1 to 1.75 0 0.7 1.00The ratios of 2-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 2-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.19 to 1/3.5.

TABLE 28 First Component (A) = 3-n-propylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.6 0 1.00 0.5 0.025 0.87 1 to 0.05 0.3 0.050.57 1 to 0.17 0.4 0.05 0.74 1 to 0.13 0.5 0.05 0.90 1 to 0.1 0.3 0.0750.61 1 to 0.25 0.4 0.075 0.77 1 to 0.19 0.5 0.075 0.94 1 to 0.15 0.2 0.10.48 1 to 0.5 0.3 0.1 0.64 1 to 0.33 0.4 0.1 0.81 1 to 0.25 0.5 0.1 0.981 to 0.2 0.08 0.35 0.63 1 to 4.38 0.1 0.35 0.67 1 to 3.5 0.2 0.35 0.83 1to 1.75 0 0.7 1.00The ratios of 3-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 3-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.05 to 1/4.4.

TABLE 29 First Component (A) = 4-n-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.2 0 1.00 0.1 0.1 0.64 1 to 1 0.03 0.35 0.651 to 11.7 0.04 0.35 0.70 1 to 8.75 0.05 0.35 0.75 1 to 7 0.06 0.35 0.801 to 5.8 0.08 0.35 0.90 1 to 4.4 0 0.7 1.00The ratios of 4-n-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 4-n-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/1 to 1/1.7.

TABLE 30 First Component (A) = 4-sec-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.2 0 1.00 0.05 0.35 0.75 1 to 7 0.06 0.350.80 1 to 5.8 0.08 0.35 0.90 1 to 4.4 0 0.7 1.00The ratios of 4-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 4-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/4.4 to 1/7.

TABLE 31 First Component (A) = 3-tert-butylphenol Second Component (B) =2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.3 0 1.00 0.2 0.075 0.77 1 to 0.38 0.2 0.10.81 1 to 0.5 0.06 0.35 0.70 1 to 5.8 0.08 0.35 0.77 1 to 4.4 0.1 0.350.83 1 to 3.5 0 0.7 1.00The ratios of 3-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested ranged from 1/0.025 to1/450. The synergistic ratios of 3-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol range from 1/0.38 to 1/5.8.

TABLE 32 First Component (A) = 2-n-propylphenol Second Component (B) =(1S,2S,5R)-2-isopropyl-5- methylcyclohexyl acetate Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.5 0 1.00 0.3 0.5 0.81 1 to 1.7 02.4 1.00The ratios of 2-n-propylphenol to(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate tested ranged from1/0.025 to 1/1100. The synergistic ratio of 2-n-propylphenol to(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate was 1/1.7.

TABLE 33 First Component (A) = 2-n-propylphenol Second Component (B) =2-(4-methylcyclohex-3- enyl)propan-2-yl butyrate Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.8 0 1.00 0.5 0.5 0.83 1 to 1 0.5 11.04 1 to 2 0.6 1.25 1.27 1 to 2.08 0.6 1.5 1.38 1 to 2.5 0.6 1.75 1.481 to 2.92 0.6 2 1.58 1 to 3.3 0.6 2.2 1.67 1 to 3.7 0 2.4 1.00The ratios of 2-n-propylphenol to 2-(4-methylcyclohex-3-enyl)propan-2-ylbutyrate tested ranged from 1/0.025 to 1/1100. The synergistic ratio of2-n-propylphenol to 2-(4-methylcyclohex-3-enyl)propan-2-yl butyrate was1/1.

TABLE 34 First Component (A) = 4-n-butylphenol Second Component (B) =(1S,2S,5R)-2-isopropyl-5- methylcyclohexyl acetate Microorganism Q_(a)Q_(b) SI Ratio A to B E. coli 10536 0.3 0 1.00 0.2 0.5 0.88 1 to 2.5 0.21 1.08 1 to 5 0 2.4 1.00The ratios of 4-n-butylphenol to(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate tested ranged from1/0.025 to 1/1100. The synergistic ratio of 4-n-butylphenol to(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate was 1/2.5.

TABLE 35 First Component (A) = 4-propylphenol Second Component (B) =2-(trans-4-ethylcyclohexyl)propane-1,3-diol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.5 0 1.00 0.3 0.025 0.68 1 to 0.08 0.4 0.0250.88 1 to 0.06 0.4 0.05 0.97 1 to 0.13 0.2 0.075 0.65 1 to 0.4 0.3 0.0750.85 1 to 0.25 0 0.3 1.00The ratios of 4-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol tested ranged from 1/0.025to 1/400. The synergistic ratios of 4-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol range from 1/0.06 to 1/0.4.

TABLE 36 First Component (A) = 2-n-propylphenol Second Component (B) =2-(trans-4-ethylcyclohexyl)propane-1,3-diol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.5 0 1.00 0.3 0.05 0.68 1 to 0.17 0.4 0.050.88 1 to 0.13 0.3 0.075 0.73 1 to 0.25 0.4 0.075 0.93 1 to 0.19 0.2 0.10.57 1 to 0.5 0.3 0.1 0.77 1 to 0.33 0.4 0.1 0.97 1 to 0.25 0.1 0.3 0.701 to 3 0.2 0.3 0.90 1 to 1.5 0 0.6 1.00The ratios of 2-n-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol tested ranged from 1/0.025to 1/400. The synergistic ratios of 2-n-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol range from 1/0.13 to 1/3.

TABLE 37 First Component (A) = 3-n-propylphenol Second Component (B) =2-(trans-4-ethylcyclohexyl)propane-1,3-diol Microorganism Q_(a) Q_(b) SIRatio A to B E. coli 10536 0.5 0 1.00 0.3 0.1 0.77 1 to 0.33 0.4 0.10.97 1 to 0.25 0.1 0.3 0.70 1 to 3 0.2 0.3 0.90 1 to 1.5 0 0.6 1.00The ratios of 3-n-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol tested ranged from 1/0.025to 1/400. The synergistic ratios of 3-n-propylphenol to2-(trans-4-ethylcyclohexyl)propane-1,3-diol range from 1/0.25 to 1/3.

TABLE 38 First Component (A) = 3-n-propylphenol Second Component (B) =2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol Microorganism Q_(a) Q_(b)SI Ratio A to B E. coli 10536 0.8 0 1.00 0.3 0.5 0.58 1 to 1.7 0.4 0.50.70 1 to 1.25 0.5 0.5 0.83 1 to 1 0.6 0.5 0.95 1 to 0.8 0.6 1 1.15 1 to1.67 0.6 1.25 1.25 1 to 2 0.6 1.5 1.35 1 to 2.5 0.6 1.75 1.45 1 to 2.9 02.5 1.00The ratios of 3-n-propylphenol to2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol tested ranged from1/0.025 to 1/1100. The synergistic ratios of 3-n-propylphenol to2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol range from 1/0.8 to1/1.7.

The following microbicidal compositions were tested and were found notto be synergistic: 3-isopropyl-5-methylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested at a weight ratio of1/0.025 to 1/350; 4-isopropyl-3-methylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested at a weight ratio of1/0.025 to 1/350; 2-tert-butyl-5-methylphenol and 2-methyl-5-(prop1-en-2-yl)cyclohexanol tested at a weight ratio of 1/0.025 to 1/350;2-sec-butylphenol and 2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested ata weight ratio of 1/0.025 to 1/350; 4-n-butylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested at a weight ratio of1/0.025 to 1/350; 4-pentylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested at a weight ratio of1/0.025 to 1/350; 3-isopropyl-5-methylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested at a weightratio of 1/0.025 to 1/350; 2-tert-butyl-5-methylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested at a weightratio of 1/0.025 to 1/350; 4-pentylphenol and(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested at a weightratio of 1/0.025 to 1/350; 3-n-propylphenol and(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate tested at a weightratio of 1/0.025 to 1/1100; 2-n-propylphenol and2-(4-methylcyclohex-3-enyl)propan-2-yl propionate tested at a weightratio of 1/0.025 to 1/1100; 3-n-propylphenol and2-(4-methylcyclohex-3-enyl)propan-2-yl propionate tested at a weightratio of 1/0.025 to 1/1100; 4-n-butylphenol and2-(4-methylcyclohex-3-enyl)propan-2-yl propionate tested at a weightratio of 1/0.025 to 1/1100; 3-n-propylphenol and2-(4-methylcyclohex-3-enyl)propan-2-yl butyrate tested at a weight ratioof 1/0.025 to 1/1100; 4-n-butylphenol and2-(4-methylcyclohex-3-enyl)propan-2-yl butyrate tested at a weight ratioof 1/0.025 to 1/1100; 2-sec-butylphenol and2-(4-methylcyclohex-3-enyl)propan-1-ol tested at a weight ratio of1/0.025 to 1/250; 4-pentylphenol and2-(4-methylcyclohex-3-enyl)propan-1-ol tested at a weight ratio of1/0.025 to 1/250; 2-sec-butylphenol and2,4,6-trimethyl-3-cyclohexene-1-methanol tested at a weight ratio of1/0.025 to 1/400; 2-tert-butylphenol and 3,7-dimethylocta-1,6-dien-3-oltested at a weight ratio of 1/0.025 to 1/350; 2-sec-butylphenol and3,7-dimethylocta-1,6-dien-3-ol tested at a weight ratio of 1/0.025 to1/350; 4-sec-butylphenol and 3,7-dimethylocta-1,6-dien-3-ol tested at aweight ratio of 1/0.025 to 1/350; 4-sec-butylphenol and(E)-3,7-dimethylocta-2,6-dien-1-ol tested at a weight ratio of 1/0.025to 1/300; 4-chloro-2-isopropyl-5-methylphenol and5-methyl-2-(prop-1-en-2-yl)cyclohexanol tested at a weight ratio of1/0.025 to 1/350; 4-chloro-2-isopropyl-5-methylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol tested at a weight ratio of1/0.025 to 1/450; 4-chloro-2-isopropyl-5-methylphenol and2-(4-methylcyclohex-3-enyl)propan-1-ol tested at a weight ratio of1/0.025 to 1/250; 4-chloro-2-isopropyl-5-methylphenol and(4S)-(4-(prop-1-en-2-yl)cyclohex-1-enyl)methanol tested at a weightratio of 1/0.025 to 1/250; 4-chloro-2-isopropyl-5-methylphenol and2,4,6-trimethyl-3-cyclohexene-1-methanol tested at a weight ratio of1/0.025 to 1/400; 4-chloro-2-isopropyl-5-methylphenol and3,7-dimethylocta-1,6-dien-3-ol tested at a weight ratio of 1/0.025 to1/250; 4-chloro-2-isopropyl-5-methylphenol and(E)-3,7-dimethylocta-2,6-dien-1-ol tested at a weight ratio of 1/0.025to 1/250; 4-chloro-2-isopropyl-5-methylphenol andcis-3,7-dimethyl-2,6-octadien-1-ol tested at a weight ratio of 1/0.025to 1/350; 2-hydroxydiphenylmethane and2-((1s,4r)-4-propylcyclohexyl)propene-1,3-diol tested at a weight ratioof 1/0.025 to 1/1100; 2-hydroxydiphenylmethane and(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate tested at a weightratio of 1/0.025 to 1/1100; 2-hydroxydiphenylmethane and2-(4-methylcyclohex-3-enyl)propan-2-yl propionate tested at a weightratio of 1/0.025 to 1/1100; 2-hydroxydiphenylmethane and2-(4-methylcyclohex-3-enyl)propan-2-yl butyrate tested at a weight ratioof 1/0.025 to 1/1100; 4-hydroxydiphenylmethane and(4S)-(4-(prop-1-en-2-yl)cyclohex-1-enyl)methanol tested at a weightratio of 1/0.025 to 1/400; 5,6,7,8-tetrahydronaphthalen-1-ol and(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate tested at a weightratio of 1/0.025 to 1/1100; 5,6,7,8-tetrahydronaphthalen-2-ol and(1S,2S,5R)-2-isopropyl-5-methylcyclohexyl acetate tested at a weightratio of 1/0.025 to 1/1100; 5,6,7,8-tetrahydronaphthalen-1-ol and2-(4-methylcyclohex-3-enyl)propan-2-yl propionate tested at a weightratio of 1/0.025 to 1/1100; 5,6,7,8-tetrahydronaphthalen-2-ol and2-(4-methylcyclohex-3-enyl)propan-2-yl propionate tested at a weightratio of 1/0.025 to 1/1100; 5,6,7,8-tetrahydronaphthalen-1-ol and2-(4-methylcyclohex-3-enyl)propan-2-yl butyrate tested at a weight ratioof 1/0.025 to 1/1100; 5,6,7,8-tetrahydronaphthalen-2-ol and2-(4-methylcyclohex-3-enyl)propan-2-yl butyrate tested at a weight ratioof 1/0.025 to 1/100; 2-cyclopentylphenol and2-methyl-5-(prop-1-en-2-yl)cyclohexanol tested at a weight ratio of1/0.025 to 1/350; and 4-pentylphenol and(4S)-(4-(prop-1-en-2-yl)cyclohex-1-enyl)methanol tested at a weightratio of 1/0.025 to 1/250.

Further experiments were conducted to determine the synergisticinteractions between the two classes of actives claimed in the presentinvention. The actives considered in this set of experiments which havecommon names are:

-   2-methyl-5-(prop-1-en-2-yl)cyclohexanol: also known as    dihydrocarveol-   2-methyl-5-(1-methylethenyl)-2-cyclohexen-1-ol or    2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol: also known as carveol-   5-methyl-2-(prop-1-en-2-yl)cyclohexanol: also known as isopulegol-   5-isopropyl-2-methylphenol: also known as carvacrol

The experimental method used for the measurements is given below:

Experimental Method

Antimicrobial efficacy is tested against a representative pathogenicbacterial organism, Gram negative Escherichia coli. Concentrations ofactives are expressed in terms of the percentage weight/volume (% w/v)throughout the following Examples.

Selected materials were also tested against a representative Grampositive Staphylococcus aureus.

E. coli Bacterial Stock

An overnight culture of Escherichia coli (10536 strain) was prepared in50 ml total volume of TSB broth, grown for ca. 18 hrs at 37° C. andshaken at 150 rpm. 1 ml of this overnight E. coli culture wastransferred to 50 ml of fresh TSB broth and incubated at 37° C. at 150rpm for ca. 4 hours. This culture was separated into equal volumes andcentrifuged at 4000 rpm for 15 minutes, washed with sterile saline(0.85% NaCl), centrifuged once more and re-suspended in saline to give afinal concentration of 0.8 OD₆₂₀ equivalent to about 10⁸ cells permillilitre for this particular organism. Here, OD₆₂₀ indicates theabsorbance of a sample in a cuvette of 1.0 cm path length at awavelength of 620 nm. This bacterial stock was used for assaying againstantimicrobial actives (in triplicate).

Protocol

The following assay describes the testing of 8 materials using 6dilutions across half of a 96-well micro titre plate (MTP). Using thisapproach it is possible to assay 16 actives (without replicates) withone full dilution plate, replicating this set up in two halves of theplate columns, 1-6 and 7-12.

1M solutions of the test actives were prepared in dimethylsulphoxide(DMSO). Stock solutions of the actives at 1.11 times the desired finalconcentration were prepared by diluting the DMSO solutions in water, sothat for example a 0.89% w/v solution was prepared for a desired “intest” concentration of 0.8% w/v in order to allow for the furtherdilution of the active when the bacterial suspension is added (dilutionfrom 270 μl to 300 μl), as described below.

Aliquots (270 μl) of the materials at 1.11 times the final concentrationwere dispensed into the wells of the MTP along one column (A1-H1). ThisMTP was labelled as the “Screening plate”.

In another MTP, labelled as the “Dilution plate”, 270 μl of D/Eneutralising solution from DIFCO Composition was added to column 1. Thecomposition of the neutralising solution was as follows: pancreaticdigest of casein, 5.0 g/L; Yeast Extract, 2.5 g/L; Dextrose, 10 g/L,sodium thioglycollate, 1.0 g/L, sodium thiosulphate, 6.0 g/L; sodiumbisulphite, 2.5 g/L; Polysorbate 80, 5.0 g/L; lecithin 7.0 g/L;bromocresol purple, 0.02 g/L with a pH in the range 7.6±0.2.

270 μl of tryptone diluent solution was added to all the remaining wellsof the Dilution MTP (columns 2-6).

Bacterial stock (30 μl) was then added to the prepared 270 μl of thesolution of actives in the Screening Plate and mixed, using amultichannel pipette with 8 tips to aspirate and dispense the samevolume of bacterial stock in parallel to 8 wells in rows A-H. After acontact time of 15 seconds, the mixtures were quenched by transferring30 μl volumes of the mixtures into the 270 μl D/E neutralising solutionin the prepared dilution plate, using aspiration to mix. After exactly 5minutes in the D/E neutralising solution, 301 volumes were transferredfrom column 1 to column 2 of the Dilution MTP and mixed, beforetransferring further 30 μl volumes from column 2 into column 3. Thisprocess was repeated serially diluting the bacteria across the plate tocolumn 6.

30 μl volumes from each well in the Dilution MTP were transferred ontopre-labelled segment of Tryptone Soya Agar (TSA) plates starting fromthe lowest bacterial concentration (highest dilution, column 6) to thehighest bacterial concentration (column 1). The TSA plates were allowedto stand for ca. 2 hours so that the 30 μl inocula spots could dry andthe plates were then inverted and incubated overnight at 37° C. beforeenumerating the bacterial colonies at the labelled dilutions todetermine the effects of the actives on bacterial growth.

Calculation of Results

Mean bacterial survival numbers N_(MBS) (expressed in Log CFU/ml) areobtained by first determining the segment of the TSA plate where thenumber of bacterial colonies is countable. From the colony number inthis segment. N s is calculated by the formula:

N _(MBS)=log{N _(col)·10^(DF)·100/3}

Here, N_(col) is the colony count, and DF is the dilution factor takenfrom the MTP-well corresponding to the TSA plate segment (i.e. DF mayrange from 1 for the quench, to 6 for the highest dilution). The factor100/3 is a conversion factor from the volume of the inocula spot to onemillilitre.

Every assay test was performed in triplicate. The reported meanbacterial survival results are the average of such a triplet, the erroris the corresponding standard deviation.

Thus, a value of N_(MBS) as of about 7 corresponds to a count of about 3colonies from the fifth dilution well, i.e. with DF=5. Such a count ofabout 7 is generally observed when bacteria are exposed to non-biocidalmaterials. In case no surviving colonies are observed in any segment ofthe TSA plate, this is interpreted as complete kill and a value ofN_(MBS)=0 is reported.

The data is summarised in the Tables below:

TABLE 39 Phenolic compound Antimicrobial alcohol concentrationconcentration N_(MBS) (% w/v) (% w/v) [log CFU/ml] 0.25% 2-propylphenol0 0 0.125% 2-propylphenol 0 0 0.06% 2-propylphenol 0 7.68 0 0.4%isopulegol 0 0 0.3% Isopulegol 0 0 0.15% isopulegol 7.77 0.125%2-propylphenol 0.1% isopulegol 0 0.06% 2-propylphenol 0.1% isopulegol 0

TABLE 40 Component MBC (% w/v) 2-propylphenol 0.125 isopulegol 0.3Minimum biocidal concentrations of antimicrobial components

TABLE 41 2-propylphenol isopulegol Evidence MBC MBC of Ex. % (w/v)FBC^(a) % (w/v) FBC^(b) ΣFBC Synergy^(c) AA 0.125 0.48 0.3 0.33 0.81 YesExtent of synergistic interactions between binary compound mixtures forcompositions providing complete bacterial kill against E. coliThe data in Table 39 to 41 indicates the synergistic interaction between2-isopropyl phenol and isopulegol which are two of the preferred activesas per the present invention.

Micobiocidal Activity of Actives as Per the Invention in a ModelSurfactant Medium Test Methodology for Automated Assessment of Efficacyin Surfactant Base

In these examples, the efficacy of combinations of actives were testedin a surfactant cleansing formulation comprising 2.85% sodium cocoylglycinate and 1.85% sodium lauroamphoacetate.

This corresponds to a 50% in use dilution with water of a neatformulation containing 5.7% cocoyl glycinate and 3.7% sodiumlauroamphoacetate during hand washing.

Solutions were prepared such that the concentrations of the surfactantcomponents and test actives were 1.1× the final desired concentration inorder to allow for dilution with the bacterial inoculum in the test. Thesolutions were manually adjusted to pH 10.0 by dropwise addition ofsodium hydroxide solution, as measured with a pH meter at ambienttemperature.

The efficacy of the combinations of the present invention was determinedagainst Escherichia coli (E. coli—ATCC #10536), at a concentration ofapproximately 1×10⁸ bacteria per mL.

Tests were conducted using standard microtiter plate assays using anautomated liquid handling system. 270 μl of the surfactant test solutionwas pipetted into each well of the microtitre plate and 30 μl of thebacterial suspension was then added. After exactly 15 seconds ofbacterial exposure, a 30 μl volume of bacterial cells was withdrawn andtransferred to 270 μl of D/E quench solution. After 5 minutes in the D/Equench, the optical density (OD) was measured for each plate in turn attwo specific wavelengths (450 nm and 590 nm). These provide a dual checkof antimicrobial activity, as the OD₄₅₀ reading is specific for theyellow colour of D/E quench when bacterial growth is observed, whereasOD₅₉₀ is specific for the initial purple colour of the D/E quench whichis retained if no bacterial growth is observed. After the time zero ODmeasurements, plates were then incubated at 37° C. overnight (16 hours)before repeating the OD measurements. Delta OD values were calculated bysubtracting the OD values at 16 hours from the initial value at timezero. Bacterial growth is observed as an increase in OD₄₅₀ and adecrease in ΔOD₅₉₀. To identify antibacterially efficacious systems(those preventing appreciable bacterial growth after incubation), thefollowing threshold changes in OD readings have been adopted: if (1).OD₄₅₀ increases by less than 0.2 absorbance unit on incubation and (2).OD₅₉₀ decreases by less than 0.35 unit on incubation. Conversely, whereOD₄₅₀ increases by more than 0.2 AU and OD₅₉₀ decreases by more than0.35 unit after incubation, corresponding to a colour shift from purpleto yellow, the test system allows bacterial growth and is not deemedefficacious. Four replicate measurements in the same plate have beenmade for each test system. The number of replicate wells showing eitherbacterial growth or no growth is also readily assessed by eye byfollowing the colour change.

Dose responses for individual components and binary mixtures of activesat a fixed concentration ratio were generated by sequential dilution ofliquors with further surfactant solution to obtain a series ofendpoints.

In each case, binary mixtures were assessed in the weight to weightratio phenol to terepene alcohol of 1:2.5. In some selected cases, thecombinations were also tested at the weight ratio 1:1.

The data is summarized in the Table below Antibacterial activities ofphenols and terpene alcohols alone and in combination in modelsurfactant solution, against E. Coli

TABLE 42 ΔOD 450 nm = Δ OD 590 nm = OD₄₅₀ (time = OD₅₉₀ (time = 16hours) − 16 hours) − No. of Phenolic Antimicrobial OD₄₅₀ OD₅₉₀replicates compound alcohol (time zero) (time zero) showingconcentration concentration Standard Standard growth (% w/v) (% w/v)Mean deviation Mean deviation (out of 4) 0.2% 0 −0.52 0.01 0.62 0.03 4carvacrol 0.125% 0 −0.50 0.03 0.63 0.02 4 carvacrol 0.075% 0 −0.52 0.030.64 0.03 4 carvacrol 0 0.5% Dihydro- 0.23 0.01 0.20 0.03 0 carveol 00.3% Dihydro- −0.44 0.01 0.66 0.01 4 carveol 0 0.15% Dihydro- −0.50 0.030.63 0.04 4 carveol 0.2% 0.5% Dihydro- 0.23 0.06 0.22 0.02 0 carvacrolcarveol 0.175% 0.4375% Dihydro- 0.20 0.04 0.22 0.02 0 carvacrol carveol0.15% 0.375% Dihydro- 0.06 0.32 0.33 0.24 1 carvacrol carveol 0.125%0.3125% Dihydro- 0.18 0.05 0.21 0.02 0 carvacrol carveol 0 0.5% −0.090.41 0.42 0.26 2 Isopulegol 0 0.3% −0.49 0.01 0.61 0.02 4 Isopulegol 00.25% −0.54 0.01 0.59 0.03 4 Isopulegol 0.2% 0.5% 0.10 0.32 0.33 0.19 1carvacrol Isopulegol 0.175% 0.4375% 0.08 0.24 0.24 0.06 1 carvacrolIsopulegol 0.15% 0.375% 0.02 0.30 0.30 0.24 1 carvacrol Isopulegol 0.2%(E)-2- 0 −0.53 0.03 0.63 0.02 4 (prop-1- enyl)phenol 0.15% (E)-2- 0−0.57 0.02 0.58 0.04 4 (prop-1- enyl)phenol 0.1% (E)-2- 0 −0.60 0.020.59 0.01 4 (prop-1- enyl)phenol 0.2% (E)-2- 0.5% Dihydro- 0.23 0.020.22 0.02 0 (prop-1- carveol enyl)phenol 0.175% (E)-2- 0.4375% Dihydro-0.21 0.02 0.25 0.03 0 (prop-1- carveol enyl)phenol 0.15% (E)-2- 0.375%Dihydro- 0.18 0.01 0.19 0.01 0 (prop-1- carveol enyl)phenol 0.2% (E)-2-0.5% 0.23 0.03 0.20 0.03 0 (prop-1- Isopulegol enyl)phenol 0.175% (E)-2-0.4375% 0.25 0.02 0.22 0.01 0 (prop-1- Isopulegol enyl)phenol 0.15%(E)-2- 0.375% −0.08 0.38 0.46 0.27 2 (prop-1- Isopulegol enyl)phenol0.15% 2- 0 −1.15 0.08 0.66 0.04 4 propylphenol 0.05% 2- 0 −1.43 0.060.46 0.07 4 propylphenol 0.2% 2- 0.2% Dihydro- −0.09 0.01 0.09 0.02 0propylphenol carveol 0.15% 2- 0.15% Dihydro- −0.09 0.01 0.09 0.00 0propylphenol carveol 0.125% 2- 0.125% Dihydro- −0.09 0.01 0.11 0.01 0propylphenol carveol 0.2% 2- 0.5% Dihydro- −0.09 0.01 0.09 0.01 0propylphenol carveol 0.15% 2- 0.375% Dihydro- −0.07 0.04 0.13 0.09 0propylphenol carveol 0.2% 2- 0.5% 0.29 0.07 0.17 0.04 0 propylphenolIsopulegol 0.175% 2- 0.4375% 0.12 0.03 0.19 0.04 0 propylphenolIsopulegol 0.15% 2- 0.375% 0.12 0.04 0.14 0.04 0 propylphenol Isopulegol0 0.5% 2- −0.48 0.02 0.590 0.02 4 ((Ir,4)-4- ethylcyclohexyl)propane-1,3-diol 0 0.4% 2- −0.49 0.04 0.56 0.04 4 ((Ir,4)-4-ethylcyclohexyl) propane-1,3-diol 0 0.25% 2- −0.54 0.02 0.56 0.03 4((Ir,4)-4- ethylcyclohexyl) propane-1,3-diol 0.2% 2- 0.5% 2- −0.11 0.010.13 0.01 0 propylphenol ((Ir,4)-4- ethylcyclohexyl) propane-1,3-diol0.175% 2- 0.4375% 2- −0.10 0.01 0.13 0.01 0 propylphenol ((Ir,4)-4-ethylcyclohexyl) propane-1,3-diol 0.15% 2- 0.375% 2- −0.09 0.03 0.160.06 0 propylphenol ((Ir,4)-4- ethylcyclohexyl) propane-1,3-diol 0.2% 3-0 −0.57 0.03 0.56 0.03 4 propylphenol 0.125% 3- 0 −0.59 0.01 0.57 0.02 4propylphenol 0.075% 3- 0 −0.63 0.02 0.56 0.02 4 propylphenol 0.2% 3-0.5% Dihydro- 0.2 0.03 0.15 0.04 0 propylphenol carveol 0.175% 3-0.4375% Dihydro- 0.18 0.06 0.14 0.06 0 propylphenol carveol 0.125% 3-0.3125% Dihydro- 0.22 0.04 0.18 0.03 0 propylphenol carveol 0.2% 3- 0.5%0.23 0.02 0.17 0.01 0 propylphenol Isopulegol 0.175% 3- 0.4375% 0.230.04 0.19 0.03 0 propylphenol Isopulegol 0.125% 3- 0.3125% 0.21 0.030.15 0.02 0 propylphenol Isopulegol 0.1% 3- 0.25% −0.51 0.04 0.58 0.04 4propylphenol Isopulegol 0.2% 4- 0 −1.49 0.02 0.43 0.03 4 propylphenol0.1% 4- 0 −1.59 0.03 0.34 0.01 4 propylphenol 0.2% 4- 0.5% Dihydro-−0.09 0.02 0.10 0.01 0 propylphenol carveol 0.175% 4- 0.4375% Dihydro-−0.09 0.01 0.10 0.02 0 propylphenol carveol 0.125% 4- 0.3125% Dihydro-−0.10 0.01 0.12 0.01 0 propylphenol carveol 0.2% 2-tert- 0 −0.04 0.010.07 0.01 0 butylphenol 0.125% 2-tert- 0 −1.48 0.02 0.30 0.01 4butylphenol 0.1% 2-tert- 0 −1.55 0.02 0.23 0.01 4 butylphenol 0.05%2-tert- 0 −1.58 0.02 0.22 0.01 4 butylphenol 0.2% 2-tert- 0.5% Dihydro-−0.05 0.00 0.08 0.00 0 butylphenol carveol 0.15% 2-tert- 0.375% Dihydro-−0.04 0.01 0.09 0.01 0 butylphenol carveol 0.1% 2-tert- 0.25% Dihydro-−0.06 0.02 0.11 0.02 0 butylphenol carveol 0.2% 2-tert- 0.5% −0.05 0.010.10 0.01 0 butylphenol Isopulegol 0.15% 2-tert- 0.375% −0.04 0.0 0.110.01 0 butylphenol Isopulegol 0.1% 2-tert- 0.25% −0.08 0.07 0.14 0.05 0butylphenol Isopulegol 0.2% 3-tert- 0 −0.47 0.02 0.64 0.02 4 butylphenol0.125% 3-tert- 0 −0.45 0.01 0.65 0.01 4 butylphenol 0.2% 3-tert- 0.5%Dihydro- 0.28 0.00 0.25 0.01 0 butylphenol carveol 0.175% 3-tert-0.4375% Dihydro- 0.27 0.06 0.25 0.05 0 butylphenol carveol 0.15% 3-tert-0.375% Dihydro- 0.25 0.02 0.25 0.03 0 butylphenol carveol 0.15% 2-sec- 0−0.41 0.29 0.50 0.30 3 butylphenol 0.1% 2-sec- 0 −1.44 0.04 0.36 0.06 4butylphenol 0.05% 2-sec- 0 −1.50 0.09 0.30 0.08 4 butylphenol 0.2%2-sec- 0.5% −0.06 0.01 0.01 0.01 0 butylphenol Isopulegol 0.15% 2-sec-0.375% −0.05 0.00 0.10 0.02 0 butylphenol Isopulegol 0.1% 2-sec- 0.25%−0.05 0.03 0.13 0.02 0 butylphenol Isopulegol 0.2% 4-sec- 0 −0.56 0.030.59 0.01 4 butylphenol 0.15% 4-sec- 0 −0.53 0.04 0.60 0.02 4butylphenol 0.125% 4-sec- 0 −0.56 0.02 0.60 0.01 4 butylphenol 0.2%4-sec- 0.5% Dihydro- 0.28 0.02 0.18 0.01 0 butylphenol carveol 0.175%4-sec- 0.4375% Dihydro- 0.13 0.03 0.18 0.05 0 butyl phenol carveol 0.2%4-sec- 0.5% 0.28 0.11 0.18 0.04 0 butyl phenol Isopulegol 0.175% 4-sec-0.4375% 0.15 0.05 0.18 0.05 0 butyl phenol Isopulegol 0.2% 4-sec- 0.5%2- −0.11 0.01 0.12 0.01 0 butyl phenol ((Ir,4)-4- ethylcyclohexyl)propane-1,3-diol 0.175% 4-sec- 0.4375% 2- −0.25 0.20 0.27 0.18 1 butylphenol ((Ir,4)-4- ethylcyclohexyl) propane-1,3-diol 0.2% 4- 0 −0.54 0.020.58 0.03 4 butylphenol 0.15% 4- 0 −0.61 0.01 0.54 0.01 4 butylphenol0.1% 4- 0 −0.61 0.01 0.53 0.02 4 butylphenol 0.2% 4- 0.5% Dihydro- 0.190.01 0.20 0.02 0 butylphenol carveol 0.175% 4- 0.4375% Dihydro- 0.190.01 0.19 0.02 0 butylphenol carveol 0.2% 4- 0.5% 0.18 0.01 0.19 0.01 0butylphenol Isopulegol 0.175% 4- 0.4375% 0.17 0.02 0.20 0.01 0butylphenol Isopulegol

In the table below the synergistic interaction of carveol with certainphenolic compounds is demonstrated. The method used was the same as inthe above Table.

TABLE 43 Δ OD 450 nm = Δ OD 590 nm = OD₄₅₀ (time = OD₅₉₀ (time = 16hours) − 16 hours) − No. of Phenolic Antimicrobial OD₄₅₀ OD₅₉₀replicates compound alcohol (time zero) (time zero) showingconcentration concentration Standard Standard growth (% w/v) (% w/v)Mean deviation Mean deviation (out of 4) 0 0.4% −0.35 0.12 0.47 0.15 4Carveol 0 0.3% −0.50 0.02 0.59 0.02 4 Carveol 0 0.2% −0.51 0.03 0.610.02 4 Carveol 0 0.15% −0.50 0.03 0.63 0.03 4 Carveol 0.2% 0.5% 0.210.05 0.21 0.02 0 carvacrol Carveol 0.175% 0.4375% 0.23 0.04 0.20 0.06 0carvacrol Carveol 0.15% 0.375% 0.17 0.04 0.19 0.02 0 carvacrol Carveol0.125% 0.3125% 0.02 0.31 0.29 0.22 1 carvacrol Carveol 0.2% 4- 0 −1.410.03 0.36 0.04 4 isopropyl-3- methyl- phenol 0.1% 4- 0 −1.50 0.02 0.240.01 4 isopropyl-3- methyl- phenol 0.05% 4- 0 −1.52 0.01 0.25 0.01 4isopropyl-3- methyl- phenol 0.175% 4- 0.4375% −0.05 0.01 0.06 0.01 0isopropyl-3- Carveol methyl- phenol 0.125% 4- 0.3125% −0.05 0.01 0.070.01 0 isopropyl-3- Carveol methyl- phenol 0.075% 4- 0.1875% −0.07 0.010.10 0.01 0 isopropyl-3- Carveol methyl- phenol 0.2% (E)-2- 0.5% 0.240.01 0.21 0.01 0 (prop-1- Carveol enyl)phenol 0.175% (E)-2- 0.4375% 0.230.03 0.22 0.03 0 (prop-1- Carveol enyl)phenol 0.15% (E)-2- 0.375% 0.210.01 0.20 0.02 0 (prop-1- Carveol enyl)phenol 0.2% 2- 0.2% −0.05 0.050.12 0.06 0 propylphenol Carveol 0.15% 2- 0.15% −0.07 0.01 0.09 0.01 0propylphenol Carveol 0.125% 2- 0.125% −0.06 0.06 0.13 0.07 0propylphenol Carveol 0.15% 2- 0.375% −0.06 0.03 0.14 0.06 0 propylphenolCarveol 0.125% 2- 0.3125% −0.08 0.01 0.11 0.02 0 propylphenol Carveol0.2% 3- 0.5% 0.23 0.02 0.18 0.01 0 propylphenol Carveol 0.15% 3- 0.375%0.17 0.05 0.15 0.06 0 propylphenol Carveol 0.125% 3- 0.3125% 0.17 0.010.15 0.05 0 propylphenol Carveol 0.2% 4- 0.5% −0.07 0.02 0.08 0.01 0propylphenol Carveol 0.15% 4- 0.375% −0.08 0.01 0.08 0.01 0 propylphenolCarveol 0.125% 4- 0.3125% −0.02 0.07 0.15 0.08 0 propylphenol Carveol0.2% 2-tert- 0.5% −0.03 0.02 0.08 0.02 0 butylphenol Carveol 0.125%2-tert- 0.3125% −0.03 0.02 0.09 0.01 0 butylphenol Carveol 0.1% 2-tert-0.25% −0.03 0.02 0.10 0.01 0 butylphenol Carveol 0.2% 3-tert- 0.5% 0.310.01 0.26 0.01 0 butylphenol Carveol 0.175% 3-tert- 0.4375% 0.28 0.010.24 0.05 0 butylphenol Carveol 0.2% 2-sec- 0.5% −0.05 0.01 0.08 0.02 0butylphenol Carveol 0.125% 2-sec- 0.3125% −0.03 0.02 0.10 0.00 0butylphenol Carveol 0.075% 2-sec- 0.1875% −0.03 0.02 0.12 0.02 0butylphenol Carveol 0.2% 4-sec- 0.5% 0.24 0.03 0.18 0.03 0 butylphenolCarveol 0.175% 4-sec- 0.4375% 0.15 0.05 0.18 0.06 0 butyl phenol Carveol0.2% 4- 0.5% 0.19 0.02 0.20 0.02 0 butylphenol Carveol 0.175% 4- 0.4375%0.18 0.02 0.22 0.03 0 butylphenol Carveol 0.15% 4- 0.375% 0.17 0.01 0.190.01 0 butylphenol Carveol

TABLE 44 Component MBC (% w/v) Dihydrocarveol 0.4 Carveol 0.4Isopulegol >0.5 Carvacrol >0.2 4-isopropyl-3-methylphenol >0.2(E)-2-(prop-1-enyl)phenol >0.2 2-propylphenol 0.175 3-propylphenol >0.24-propylphenol >0.2 2-tert-butylphenol 0.2 3-tert-butylphenol >0.22-sec-butylphenol 0.175 4-sec-butylphenol >0.2 4-butylphenol >0.22-((1r,4)-4-ethylcyclohexyl)propane-1,3-diol >0.2The minimum biocidal concentrations of antimicrobial components in 2.85%sodium cocoyl glycinate+1.85% sodium lauroamphoacetate solution at pH 10is given in the table below.Antimicrobial Efficacy of Actives Against S. aureus in Water

S. aureus Bacterial stock was prepared as shown below and theantibacterial efficacy of the active was carried out as explainedearlier for E. Coli.

An overnight culture of Staphylococcus aureus (10788 strain) wasinoculated onto a Tripticase Soy Agar (TSA) plate and grown for ca. 18hrs at 37° C. The culture of S. aureus was washed from the plate usingsterile saline (0.85% NaCl) solution and a sterile spreader andsuspended in 50 ml saline. This suspension was separated into equalvolumes and centrifuged at 4000 rpm for 15 minutes. Followingcentrifugation the pellet was re-suspended in saline to give a finalconcentration of 0.35 OD₆₂₀ equivalent to about 10⁸ cells per millilitrefor this particular organism. Here. OD₆₂₀ indicates the absorbance of asample in a cuvette of 1.0 cm path length at a wavelength of 620 nm.This bacterial stock was used for assaying against antimicrobial actives(in triplicate).

TABLE 45 Phenolic compound Antimicrobial N_(MBS) concentration alcoholconcentration [log Standard (% w/v) (% w/v) CFU/ml] deviation 0.25% 2- 00 0 propylphenol 0.125% 2- 0 7.19 0.06 propylphenol 0.03% 2- 0 7.12 0.21propylphenol 0 1.5% carveol 7.16 0.19 0 0.75% carveol 7.22 0.21 0 0.5%carveol 7.27 0.25 0.5% 2- 0.5% carveol 0.00 0.00 propylphenol 0.25% 2-0.25% carveol 0.00 0.00 propylphenol0 0.125% 2- 0.125% carveol 4.98 0.04propylphenol 0.05% 2- 0.05% carveol 7.07 0.13 propylphenol 0 1.5%dihydrocarveol 7.22 0.27 0 0.75% dihydrocarveol 7.39 0.15 0 0.5%dihydrocarveol 7.34 0.07 0.5% 2- 0.5% dihydrocarveol 0.00 0.00propylphenol 0.25% 2- 0 25% dihydrocarveol 0.00 0.00 propylphenol 0.125%2- 0.125% dihydrocarveol 4.68 0.05 propylphenolAntibacterial activities of 2-propylphenol alone and in combination withdihydrocarveol or carveol against S. Aureus was measured and the data issummarised below:The data in Table 44 above demonstrates the synergistic interactionbetween the actives against a highly resistant bacteria S. Aureus.

TABLE 46 Component MBC (% w/v) 2-propylphenol 0.25 Dihydrocarveol >1.5Minimum biocidal concentrations of antimicrobial components against S.aureusComplete bacterial kill is achieved by 0.25% 2-propylphenol alone and incombination with 0.25% dihydrocarveol. Although complete kill is notachieved at lower phenol concentrations, it is clear that there is anenhancement in bactericidal performance through addition of 0.125%dihydrocarveol to 0.125% 2-propylphenol.Synergy of Terpinyl Alcohols with Phenolic Compounds in Water Against S.aureus,

The procedure used was as follows:

Actives:

Liquid actives were used directly. Solid actives were dissolved inalcohol and a 50% stock was made.

Preparation of test cultures:

The culture was grown overnight in Tryptone soya agar. It was suspendedin saline (0.85% NaCl) to a con. of 1.5×10⁸-5×10⁸ cells/ml

Assay:

To 9 ml water the actives were added to give the desired concentration.1 ml of the culture was added and mixed. After 15 seconds contact 1 mlaliquot was removed and added to a neutraliser and kept for 5 minutes. Acontrol of untreated cells was similarly processed. It was then dilutedand plated. The plates were incubated at 37° C. for 24 hours. Thecolonies on the plates were counted. The counts were converted to log₁₀.The log .no. of survivors was determined and log reduction calculated.The data is summarized in the Table below.

TABLE 47 Log reduction S. aureus Phenolic compound Antimicrobial alcoholATCC 6538 0.1% 2- 1.92 propenylphenol 0.1% 2- 0.25% Dihydrocarveol 3.55propenylphenol 0.25% dihydrocarveol −0.15

1. A synergistic microbicidal composition comprising: (a) at least onemicrobicide selected from the group consisting of2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol,2-methyl-5-(prop-1-en-2-yl)cyclohexanol and5-methyl-2-(prop-1-en-2-yl)cyclohexanol; and (b) at least onemicrobicide selected from the group consisting of5-isopropyl-2-methylphenol; 3-isopropyl-5-methylphenol,4-isopropyl-3-methylphenol, (E)-2-(prop-1-enyl)phenol, 4-propylphenol,2-tert-butylphenol, 2-sec-butylphenol, 2-n-propylphenol,3-n-propylphenol, 4-n-butylphenol, 4-pentylphenol, 4-sec-butylphenol,and 3-tert-butylphenol.
 2. A synergistic microbicidal composition ofclaim 1 comprising: (a) at least one microbicide selected from the groupconsisting of 2-methyl-5-(prop-1-en-2-yl)cyclohexanol and5-methyl-2-(prop-1-en-2-yl)cyclohexanol; and (b) at least onemicrobicide selected from the group consisting of5-isopropyl-2-methylphenol, 4-isopropyl-3-methylphenol,(E)-2-(prop-1-enyl)phenol 4-propylphenol, 2-tert-butylphenol,2-sec-butylphenol, 2-n-propylphenol, 3-n-propylphenol, 4-n-butylphenol,4-sec-butylphenol, and 3-tert-butylphenol.
 3. A synergistic microbicidalcomposition of claim 1 comprising: (a) at least one microbicide selectedfrom the group consisting of 2-methyl-5-(prop-1-en-2-yl)cyclohexanol and5-methyl-2-(prop-1-en-2-yl)cyclohexanol; and (b) at least onemicrobicide selected from the group consisting of 2-n-propylphenol,4-n-butylphenol, and 4-sec-butylphenol.
 4. The synergistic microbicidalcomposition of claim 1 comprising: (a)2-methyl-5-(prop-1-en-2-yl)cyclohexanol; and (b) at least onemicrobicide selected from the group consisting of(E)-2-(prop-1-enyl)phenol, 4-propylphenol, 2-n-propylphenol,3-n-propylphenol, 4-n-butylphenol, 4-sec-butylphenol, and3-tert-butylphenol.
 5. The synergistic microbicidal composition of claim4 in which a weight ratio of (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/0.06 to 1/2.5; aweight ratio of 4-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/0.08 to 1/5; a weightratio of 2-n-propylphenol to 2-methyl-5-(prop-1-en-2-yl)cyclohexanol isfrom 1/0.05 to 1/3.13; a weight ratio of 3-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/0.08 to 1/2.5; aweight ratio of 4-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/1 to 1/3.13; and aweight ratio of 3-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohexanol is from 1/0.38 to 1/3.13.
 6. Thesynergistic microbicidal composition of claim 1 comprising: (a)(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol; and (b) at least onemicrobicide selected from the group consisting of5-isopropyl-2-methylphenol, 4-isopropyl-3-methylphenol,(E)-2-(prop-1-enyl)phenol, 4-propylphenol, 2-tert-butylphenol,2-sec-butylphenol, 2-n-propylphenol, 3-n-propylphenol, 4-n-butylphenol,and 3-tert-butylphenol.
 7. The synergistic microbicidal composition ofclaim 6 in which a weight ratio of 5-isopropyl-2-methylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.06 to1/2.5; a weight ratio of 4-isopropyl-3-methylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.5 to1/2.5; a weight ratio of (E)-2-(prop-1-enyl)phenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.13 to1/1.25; a weight ratio of 4-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.5 to1/2.5; a weight ratio of 2-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.17 to1/2.5; a weight ratio of 2-sec-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.5 to1/2.5; a weight ratio of 2-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.05 to1/1.25; a weight ratio of 3-n-propylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.06 to1/2.5; a weight ratio of 4-n-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.13 to1/2.5; and a weight ratio of 3-tert-butylphenol to(1R,2S,5R)-5-methyl-2-(prop-1-en-2-yl)cyclohexanol is from 1/0.13 to1/0.5.
 8. A synergistic microbicidal composition comprising: (a)2-(trans-4-ethylcyclohexyl)propane-1,3-diol or2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol; and (b) at least onemicrobicide selected from the group consisting of 4-propylphenol,2-n-propylphenol and 3-n-propylphenol.
 9. A synergistic microbicidalcomposition of claim 8 comprising: (a)2-(trans-4-ethylcyclohexyl)propane-1,3-diol; and (b) at least onemicrobicide selected from the group consisting of 4-propylphenol,2-n-propylphenol and 3-n-propylphenol.
 10. A synergistic microbicidalcomposition of claim 8 comprising: (a)2-((1S,4R)-4-propylcyclohexyl)propane-1,3-diol; and (b)3-n-propylphenol.
 11. A synergistic microbicidal composition of claim 1comprising: (a) 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol; and (b) atleast one microbicide selected from the group consisting of5-isopropyl-2-methylphenol; 3-isopropyl-5-methylphenol,4-isopropyl-3-methylphenol, (E)-2-(prop-1-enyl)phenol, 4-propylphenol,2-tert-butylphenol, 2-sec-butylphenol, 2-n-propylphenol,3-n-propylphenol, 4-n-butylphenol, 4-pentylphenol, 4-sec-butylphenol,and 3-tert-butylphenol.
 12. A synergistic antimicrobial composition ofclaim 11 comprising (a) 2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol and(b) at least one microbicide selected from the group consisting of2-n-propylphenol, 4-n-butylphenol, and 4-sec-butylphenol.
 13. Thesynergistic microbicidal composition of claim 12 in which a weight ratioof 3-isopropyl-5-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.38 to 1/4.38, aweight ratio of 4-isopropyl-3-methylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.25 to 1/5.8, aweight ratio of (E)-2-(prop-1-enyl)phenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.17 to 1/1.75, aweight ratio of 4-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.08 to 1/4.4, aweight ratio of 2-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.38 to 1/4.38, aweight ratio of 2-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.38 to 1/4.38, aweight ratio of 2-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.19 to 1/3.5, aweight ratio of 3-n-propylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.05 to 1/4.38, aweight ratio of 4-n-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/1 to 1/11.7, aweight ratio of 4-sec-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/4.4 to 1/7, and aweight ratio of 3-tert-butylphenol to2-methyl-5-(prop-1-en-2-yl)cyclohex-2-enol is from 1/0.38 to 1/5.8. 14.The synergistic microbicidal composition according to any one of thepreceding claims comprising from 1 to 80% by weight of one or moresurfactants.
 15. A method of disinfecting a surface comprising the stepsof a. applying a composition according to any one of the precedingclaims on to the surface; and b. removing the composition from thesurface.